CN109749096B - Carbon nanotube modified gelatin hydrogel and preparation method and application thereof - Google Patents
Carbon nanotube modified gelatin hydrogel and preparation method and application thereof Download PDFInfo
- Publication number
- CN109749096B CN109749096B CN201811578203.3A CN201811578203A CN109749096B CN 109749096 B CN109749096 B CN 109749096B CN 201811578203 A CN201811578203 A CN 201811578203A CN 109749096 B CN109749096 B CN 109749096B
- Authority
- CN
- China
- Prior art keywords
- carbon nanotube
- modified
- gelatin
- substrate
- carbon
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Abstract
The invention relates to a gelatin hydrogel modified by carbon nano tubes and a preparation method and application thereof. The preparation method of the carbon nanotube modified gelatin hydrogel comprises the following steps: carrying out homopolymerization on a methacrylate monomer to obtain a methacrylate polymer; carrying out grafting reaction on the carbon nanotube array and a methacrylate polymer to obtain a modified carbon nanotube array; reacting the modified carbon nanotube array with gelatin to obtain a carbon nanotube modified gelatin prepolymer; and crosslinking the gelatin prepolymer modified by the carbon nano tube to obtain the gelatin hydrogel modified by the carbon nano tube. The carbon nanotubes used in the carbon nanotube modified gelatin hydrogel are modified by methacrylate polymers, are easy to disperse, and can be combined with gelatin through covalent bonds and crosslinked to obtain the carbon nanotube modified gelatin hydrogel with better mechanical properties and less influence on porosity.
Description
Technical Field
The invention relates to the field of hydrogel, in particular to carbon nanotube modified gelatin hydrogel and a preparation method and application thereof.
Background
The hydrogel can be used as a three-dimensional bionic extracellular matrix support due to the characteristics of porous structure, higher water content, controllable biodegradability and the like, and provides mechanical support for cell growth and tissue formation.
In general, hydrogels as three-dimensional biomimetic extracellular matrix scaffolds need to have sufficient mechanical strength to match the tissue to be simulated, such as brain, muscle or bone, where the mechanical strength can be controlled by parameters such as density, cross-linking density or molecular weight. However, highly crosslinked three-dimensional hydrogels, although having a high stiffness, can hinder cell proliferation, migration, and morphogenesis. The pre-polymerization solution with high polymer concentration can improve the mechanical properties, but the degradability, porosity, and cell diffusion and growth are affected. Therefore, an ideal hydrogel should have its mechanical properties increased without reducing its porosity or other beneficial properties.
To meet these requirements, carbon nanotubes have been added to enhance the tensile modulus and stiffness of hydrogels. However, it is difficult to uniformly disperse the carbon nanotubes due to the property of the carbon nanotubes of being agglomerated. Meanwhile, in the traditional technology, the interaction between the carbon nanotubes and the hydrogel is weak, so that the mechanical properties of the obtained gelatin hydrogel modified by the carbon nanotubes can not meet the requirements.
Disclosure of Invention
Accordingly, there is a need for a method for preparing a gelatin hydrogel modified with carbon nanotubes having good mechanical properties and less influence on porosity.
In addition, a gelatin hydrogel modified by the carbon nano tube and application of the gelatin hydrogel modified by the carbon nano tube are also provided.
A preparation method of gelatin hydrogel modified by carbon nano tubes is characterized by comprising the following steps:
homopolymerization reaction is carried out on methacrylate monomer to obtain the product with the structural formula
The methacrylate polymer of (1), wherein n is 1 to 11, and R ═ CmH2m+1M is 1 to 4;
carrying out grafting reaction on the carbon nanotube array and the methacrylate polymer to obtain a modified carbon nanotube array;
reacting the modified carbon nanotube array with gelatin to obtain a carbon nanotube modified gelatin prepolymer; and
and crosslinking the carbon nanotube modified gelatin prepolymer to obtain the carbon nanotube modified gelatin hydrogel.
According to the preparation method of the gelatin hydrogel modified by the carbon nano tubes, the carbon nano tube array is modified by the methacrylate polymer with the unsaturated group terminal, so that the agglomeration caused by the van der Waals force between the carbon nano tubes can be reduced. The ester group on the modified carbon nanotube array is connected with the amino or hydroxyl in the gelatin through a covalent bond, and the unsaturated bond in the methacrylate polymer grafted on the carbon nanotube is crosslinked, so that the carbon nanotube modified gelatin hydrogel with better mechanical property is obtained, and experiments prove that the influence on the porosity of the hydrogel caused by adding the carbon nanotube modification is small.
In one embodiment, the step of performing homopolymerization on a methacrylate monomer to obtain a methacrylate polymer specifically includes: and carrying out homopolymerization on the methacrylate monomer, the catalyst, the initiator and the solvent under the condition of constant-temperature water bath to obtain the methacrylate polymer.
In one embodiment, the step of performing a grafting reaction between the carbon nanotube array and the methacrylate polymer specifically includes:
preparing the carbon nanotube array on a first substrate;
depositing the methacrylate-based polymer on a second substrate; and
and simultaneously carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate so as to carry out grafting reaction on the methacrylate polymer and the carbon nanotube array.
In one embodiment, the step of reacting the modified carbon nanotube array with gelatin specifically comprises:
dissolving the gelatin in phosphate buffered saline to obtain a gelatin solution;
adding the modified carbon nanotube array into the gelatin solution to obtain a mixture; and
and dialyzing the mixture, and drying to obtain the carbon nanotube modified gelatin prepolymer.
In one embodiment, the mass ratio of the modified carbon nanotube array to the gelatin is 1: 9-1: 19.
In one embodiment, the step of crosslinking the carbon nanotube modified gelatin prepolymer specifically comprises:
dissolving the carbon nanotube modified gelatin prepolymer in phosphate buffered saline to obtain a carbon nanotube modified gelatin prepolymer solution; and
adding a photoinitiator into the carbon nanotube modified gelatin prepolymer solution, and crosslinking under the irradiation of ultraviolet light B.
In one embodiment, the mass ratio of the photoinitiator to the carbon nanotube modified gelatin prepolymer is 5-8: 100.
In one embodiment, the ultraviolet light B is monochromatic narrow-band light with the wavelength of 365-400 nm, the irradiation power of the ultraviolet light B is 10-15 mW, and the processing time of the ultraviolet light B is 30-60 min.
The carbon nano tube modified gelatin hydrogel prepared by the preparation method.
An application of gelatin hydrogel modified by carbon nano tube in preparing three-dimensional cell culture support or preparing bionic extracellular matrix.
Drawings
FIG. 1 is a flow chart of a method for preparing a carbon nanotube-modified gelatin hydrogel according to an embodiment;
fig. 2 is a schematic view of the modified carbon nanotubes obtained in step S120 of the method for preparing a carbon nanotube-modified gelatin hydrogel shown in fig. 1;
fig. 3 is a schematic view of the carbon nanotube-modified gelatin prepolymer obtained in step S130 in the method for preparing a carbon nanotube-modified gelatin hydrogel shown in fig. 1;
fig. 4 is a schematic view illustrating the preparation of the carbon nanotube-modified gelatin hydrogel obtained in step S140 in the method for preparing a carbon nanotube-modified gelatin hydrogel shown in fig. 1;
FIG. 5-a is a peak separation chart of C1s in the XPS spectrum of the pure carbon nanotube in example 5; FIG. 5-b is a graph of the XPS spectrum of O1s for pure carbon nanotubes from example 5;
FIG. 6-a is a peak separation chart of C1s in the XPS spectrum of pure polymethyl methacrylate in example 5; FIG. 6-b is a peak separation chart of O1s in the XPS spectrum of pure polymethyl methacrylate in example 5;
FIG. 7-a is a peak separation plot of C1s in the XPS spectrum of the modified carbon nanotube array of example 5; FIG. 7-b is a peak separation plot of O1s in the XPS spectrum of the modified carbon nanotube array of example 5;
fig. 8 is a stress-strain diagram of the carbon nanotube-modified gelatin hydrogel of examples 1 to 5 and the methyl methacrylate-modified gelatin hydrogel of comparative example 1.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Referring to fig. 1, according to the preparation method of the carbon nanotube-modified gelatin hydrogel of an embodiment, the carbon nanotube-modified gelatin hydrogel having a good mechanical property and a small influence on the porosity of the hydrogel can be obtained.
The method for preparing the carbon nanotube-modified gelatin hydrogel of an embodiment includes the following steps:
s110: homopolymerization reaction is carried out on methacrylate monomer to obtain the product with the structural formula
The methacrylic acid ester-based polymer of (1).
Wherein n is 1-11, and R ═ CmH2m+1And m is 1 to 4. Specifically, the polymerization degree of the methacrylate polymer is 2-12, the molecular weight is 200-1200, and the polydispersity index (Polyd)ispersity Index) of 1.10 to 1.20.
Specifically, the step of homopolymerizing a methacrylate monomer to obtain a methacrylate polymer comprises:
s111: methacrylate monomer, catalyst, initiator and solvent are added separately to the reaction vessel and sealed.
Specifically, the methacrylate monomer is selected from one of methyl methacrylate, ethyl methacrylate, n-propyl methacrylate and n-butyl methacrylate. The catalyst is a divalent cobalt chelate, and the mass ratio of the catalyst to the monomer is 4.0-4.2: 100. Further, the catalyst is tetramethoxyphenyl cobalt porphyrin, cobalt tetra (tert-butyl) phthalocyanine or cobalt bis (borodifluorodiphenylglyoxime) dihydrate.
Specifically, the initiator is an azo compound, and the mass ratio of the initiator to the monomer is 2.0-2.2: 100. Further, the initiator is azobisisobutyronitrile, azobisisoheptonitrile or dimethyl azobisisobutyrate. The initiator is capable of generating carbon-centered radicals without destroying the metal chelate catalyst, which facilitates the production of oligomers.
Specifically, the solvent is chloroform, and the volume ratio of the solvent to the monomer is 6.5-7.5: 1. Further, the method further comprises the step of degassing the reaction vessel before sealing the reaction vessel.
S112: the reaction vessel was immersed in a constant temperature water bath for reaction.
Specifically, the temperature of the constant-temperature water bath is 70-80 ℃, and the reaction time is 1 h.
S113: and after the reaction is finished, removing the catalyst and the solvent to obtain the methacrylate polymer.
Specifically, the catalyst is precipitated with a mixture of acetone and water or pure ethyl acetone, and thus removed by filtration. The reagent can effectively deposit the metal cobalt chelate. The above-mentioned method for removing the solvent may be a rotary evaporation method.
Further, the methyl methacrylate polymer has an unsaturated double bond at the terminal, and polymerization can be continued.
S120: and carrying out grafting reaction on the carbon nano tube array by using a methacrylate polymer to obtain the modified carbon nano tube array. Please refer to fig. 2, which shows a schematic diagram of the modified carbon nanotube, wherein n is 1 to 11.
Specifically, the step of performing a grafting reaction on the carbon nanotube array by using a methacrylate polymer comprises:
s121: a carbon nanotube array is prepared on a first substrate.
Specifically, a carbon nanotube array is prepared by a chemical vapor deposition method. Further, the step of preparing the carbon nanotube array includes: depositing a catalyst layer on the first substrate by adopting an electron beam evaporation method, then heating the first substrate to 550-900 ℃ in a protective gas atmosphere to ensure that the catalyst layer uniformly nucleates on the first substrate, and then introducing a carbon source gas for reaction to obtain the carbon nano tube array.
Further, the catalyst layer is a cobalt-nickel alloy layer. The carbon source gas comprises 25-40% of ethylene, 1-10% of hydrogen and the balance of nitrogen (calculated by gas partial pressure). The flow rate of the introduced carbon source gas is 1L/min-3L/min, and the time for introducing the carbon source gas to react is 2 min-5 min.
Further, the first substrate is a nickel sheet or a copper sheet. The first substrate is mainly used for bearing the carbon nanotube array, and the nickel sheet and the copper sheet have good stability on the carbon nanotube array and cannot react with the carbon nanotube array. In one embodiment, the first substrate has a diameter of 8 feet. It is understood that in other embodiments, the size of the first substrate may be any other size.
Specifically, the protective gas is selected from at least one of nitrogen, argon, and helium. The introduction of the protective gas can prevent the carbon source gas from being oxidized at high temperature.
In one embodiment, the carbon nanotube array deposited on the first substrate is a single-walled carbon nanotube array. It is understood that in other embodiments, the carbon nanotube array deposited on the first substrate may be a multi-walled carbon nanotube array.
In one embodiment, the length of the carbon nanotube array deposited on the first substrate is 0.5 μm to 2.0 μm, and the diameter of the carbon nanotubes in the carbon nanotube array is 10nm to 15 nm. It is understood that the length of the carbon nanotube array may be other values in other embodiments.
S122: depositing a methacrylate polymer on the second substrate.
Specifically, the second substrate is a nickel sheet or a copper sheet. The second substrate is mainly used for bearing the methacrylate polymer, and the nickel sheet and the copper sheet have good stability and cannot react with the methacrylate polymer. In one embodiment, the second substrate has dimensions of 50mm by 50 mm. It is understood that in other embodiments, the size of the second substrate may be any other size.
Further, the methacrylate-based polymer was deposited on the second substrate in the form of a thin film, and the thickness of the thin film was 1 mm.
S123: and simultaneously, carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate so as to carry out grafting reaction on the methacrylate polymer and the carbon nanotube array to obtain the modified carbon nanotube array.
Specifically, the first substrate and the second substrate are on the same horizontal plane and are placed side by side. The ultraviolet light a irradiation treatment is performed in a protective gas atmosphere. The irradiation power of the ultraviolet light A is 15 mW-35 mW. Under the irradiation power, the formation of the gaseous methacrylate polymer can be promoted, and the gaseous methacrylate polymer is moved to the surface of the carbon nanotube array under the action of the protective gas flow, so that the grafting reaction is carried out with the carbon nanotube array.
Further, the irradiation wavelength of the ultraviolet light A is monochromatic narrow-band light of 196nm to 350 nm. Under the irradiation wavelength, carbon-carbon bonds can be effectively opened on the surface of the carbon nano tube to form dangling bonds, and meanwhile, the damage of ultraviolet light to the methacrylate polymer and the carbon nano tube array structure is reduced under the condition that the methacrylate polymer can be grafted to the carbon nano tube array. The time of the ultraviolet A treatment is 20min to 50min, and the distance between the light source of the ultraviolet A and the first substrate and the distance between the light source of the ultraviolet A and the second substrate are 2mm to 20 mm.
Further, the method also comprises the step of exposing the first substrate to the protective atmosphere until the first substrate is naturally cooled after the ultraviolet light A irradiation is finished.
The methacrylate polymer with ester group and unsaturated terminal group is grafted on the surface of the carbon nano tube, so that the distance between the carbon nano tubes can be increased, the agglomeration caused by Van der Waals force between the carbon nano tubes can be reduced, the modified carbon nano tube array is easy to disperse, and the grafted polymer has a plurality of active sites and can continue to react with other substances.
S130: and reacting the modified carbon nanotube array with gelatin to obtain the carbon nanotube modified gelatin prepolymer. Referring to fig. 3, the schematic diagram of the carbon nanotube modified gelatin prepolymer is shown, in which the methyl methacrylate polymer grafted by the carbon nanotube reacts with the amino group and the hydroxyl group on the gelatin to form a covalent bond, where n is 1 to 11.
Specifically, the step of reacting the modified carbon nanotube array with gelatin comprises:
s131: gelatin was dissolved in phosphate buffered saline to give a gelatin solution.
Further, the pH value of the phosphate buffered saline is 7-8. The dissolving temperature of the gelatin in the phosphate buffer saline is 40-50 ℃.
S132: and adding the modified carbon nanotube array into a gelatin solution for reaction to obtain a mixture.
Specifically, the mass ratio of the modified carbon nanotube array to the gelatin is 1: 9-1: 19.
Specifically, the reaction of the modified carbon nanotube array with the gelatin solution is performed under stirring conditions. The stirring time was 2 h. The stirring is conventional magnetic stirring or mechanical stirring. Thereafter, optionally, a step of adding phosphate buffered saline for dilution is taken to stop the reaction.
S133: and dialyzing the mixture, and drying to obtain the carbon nano tube modified gelatin prepolymer.
Further, the dialyzing agent is deionized water. The temperature in the dialysis process is 40-50 ℃. The dialysis time was 1 week. Small molecular substances and inorganic salts can be removed through dialysis, and furthermore, the molecular weight intercepted by a dialysis bag used in dialysis is 2000-3000.
The modified carbon nanotube array is grafted with methacrylate polymers, so that the modified carbon nanotube array has ester groups, can react with amino and hydroxyl on gelatin molecules, and is connected with gelatin through covalent bonds.
S140: and crosslinking the gelatin prepolymer modified by the carbon nano tube to obtain the gelatin hydrogel modified by the carbon nano tube. Please refer to fig. 4, wherein n is 1-11, Gelatin represents Gelatin, Initiator represents Initiator, and R represents Initiator radical.
Specifically, the step of crosslinking the carbon nanotube-modified gelatin prepolymer comprises:
s141: and dissolving the carbon nanotube modified gelatin prepolymer in phosphate buffered saline to obtain a carbon nanotube modified gelatin prepolymer solution.
Specifically, the pH of the phosphate buffered saline is 7-8. The dissolving temperature of the gelatin in the phosphate buffer saline is 70-80 ℃. Further, the above dissolution process was performed under stirring for 30 min. The stirring is conventional magnetic stirring or mechanical stirring.
S142: adding a photoinitiator into the carbon nano tube modified gelatin prepolymer solution, and crosslinking under the irradiation of ultraviolet light B.
Specifically, the mass ratio of the photoinitiator to the carbon nanotube modified gelatin prepolymer is 5-8: 100. Further, the photoinitiator was a commercially available BASF photoinitiator Irgacure 2959 or lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate. The photoinitiator is water-soluble, and therefore can be dissolved in the carbon nanotube modified gelatin prepolymer solution and can initiate crosslinking of the carbon nanotube modified gelatin prepolymer solution.
Further, the ultraviolet light B for crosslinking the gelatin prepolymer solution modified by the carbon nano tube is monochromatic narrow-band light with the wavelength of 365 nm-400 nm, and the irradiation power of the ultraviolet light B is 10 mW-15 mW. The time of the ultraviolet B treatment is 30-60 min. Furthermore, the distance between the ultraviolet light B and the gelatin prepolymer solution modified by the carbon nano tube is 20 mm-30 mm. Under the combined action of the photoinitiator and the ultraviolet light B, unsaturated double bonds on the gelatin prepolymer modified by the carbon nano tube can be subjected to free radical polymerization, so that the gelatin hydrogel modified by the carbon nano tube is obtained through crosslinking.
Specifically, the gelatin hydrogel modified by the carbon nano tube is a composite film, and the thickness of the film is 1 mm-2 mm.
In the preparation method of the gelatin hydrogel modified by the carbon nanotubes, the distance between the carbon nanotubes can be increased and the agglomeration caused by the van der waals force between the carbon nanotubes can be reduced by modifying the carbon nanotube array with the methacrylate polymer with the unsaturated group terminal. The ester group on the modified carbon nanotube array is connected with the amino or hydroxyl in the gelatin through a covalent bond, and the unsaturated bond in the methacrylate polymer grafted on the carbon nanotube is subjected to photo-crosslinking, so that the carbon nanotube modified gelatin hydrogel with better mechanical property is obtained, and experiments prove that the porosity of the hydrogel is not influenced after the carbon nanotube is added for modification. Combined with the characteristics of the hydrogel, the hydrogel can be applied to the field of biological materials such as cell scaffolds and bionic extracellular matrices.
The following are specific examples:
unless otherwise specified, the following examples contain no other components not specifically indicated except for inevitable impurities.
Example 1
(1) To a reaction vessel were added 8mg of cobalt tetramethoxyphenylporphyrin, 190mg of n-butyl methacrylate monomer, 4mg of azobisisobutyronitrile and 1.48mL of chloroform, followed by degassing treatment, and the reaction vessel was sealed. And (3) immersing the sealed reaction container into a constant-temperature water bath at 70 ℃ for reaction for 1 h. After the reaction is finished, ethyl acetone is added to precipitate the tetramethoxyphenyl porphyrin cobalt catalyst, and after filtration, the butyl methacrylate polymer is obtained by rotary evaporation. The structural formula of the n-butyl methacrylate polymer is as follows:
(2) depositing a cobalt-nickel alloy catalyst layer on a first substrate, and placing the first substrate in a chemical vapor deposition reaction furnace. Nitrogen was introduced and the temperature was raised to 550 ℃. And then introducing 25% of ethylene, 5% of hydrogen and 70% of nitrogen, and reacting for 2min to obtain the carbon nanotube array. A second substrate was taken, and an n-butyl methacrylate polymer film having a thickness of 1mm was formed on the second substrate. And in a nitrogen atmosphere, placing the first substrate and the second substrate in a reaction furnace side by side, wherein the first substrate and the second substrate are positioned on the same horizontal plane, and carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate. The irradiation power of the ultraviolet light A is 20mW, the irradiation wavelength of the ultraviolet light A is 256nm, and the irradiation time is 20 min. And after the reaction is finished, closing the ultraviolet light A, and exposing the first substrate to a nitrogen atmosphere until the first substrate is naturally cooled to obtain the modified nanotube array.
(3) Gelatin was dissolved in phosphate buffered saline at pH 7.5 at a dissolution temperature of 40 ℃. Adding a modified carbon nanotube array with the mass ratio of the modified carbon nanotube array to the gelatin being 1:19, stirring for 2 hours, and diluting with phosphate buffered saline to stop the reaction. The mixture was then dialyzed in deionized water at 40 ℃ for 1 week to remove small molecules and salts, followed by freeze-drying to give a carbon nanotube-modified gelatin prepolymer.
(4) Dissolving the gelatin prepolymer modified by the carbon nano tube in phosphate buffered saline with the pH value of 7.5, wherein the dissolving temperature is 70 ℃, fully stirring for 30min, adding a photoinitiator Irgacure 2959 with the mass ratio of the carbon nano tube modified gelatin prepolymer to the mass ratio of 5: 100, and carrying out ultraviolet B crosslinking reaction for 30min to obtain the gelatin hydrogel modified by the carbon nano tube. Wherein the irradiation power of the ultraviolet light B is 10mW, and the irradiation wavelength of the ultraviolet light B is 365 nm.
Example 2
(1) To a reaction vessel were added 8mg of cobalt tetramethoxyphenylporphyrin, 190mg of methyl methacrylate monomer, 4mg of azobisisobutyronitrile and 1.30mL of chloroform, followed by degassing treatment, and the reaction vessel was sealed. And (3) immersing the sealed reaction container into a constant-temperature water bath at 70 ℃ for reaction for 1 h. After the reaction is finished, ethyl acetone is added to precipitate the tetramethoxyphenyl porphyrin cobalt catalyst, and after filtration, the methyl methacrylate polymer is obtained by rotary evaporation. The structural formula of the methyl methacrylate polymer is as follows:
(2) depositing a cobalt-nickel alloy catalyst layer on a first substrate, and placing the first substrate in a chemical vapor deposition reaction furnace. Introducing hydrogen, and heating to 630 ℃. And then introducing 25% of ethylene, 5% of hydrogen and 70% of nitrogen at the gas flow rate of 2L/min, and reacting for 3min to obtain the carbon nanotube array. A second substrate was taken, and a methyl methacrylate polymer film having a thickness of 1mm was formed on the second substrate. And in an argon atmosphere, placing the first substrate and the second substrate in a reaction furnace side by side, wherein the first substrate and the second substrate are positioned on the same horizontal plane, and carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate. The irradiation power of the ultraviolet light A is 20mW, the irradiation wavelength of the ultraviolet light A is 216nm, and the irradiation time is 20 min. And after the reaction is finished, closing the ultraviolet light, and exposing the first substrate to a hydrogen atmosphere until the first substrate is naturally cooled to obtain the modified nanotube array.
(3) Gelatin was dissolved in phosphate buffered saline at pH 7 at a dissolution temperature of 40 ℃. Adding a modified carbon nanotube array with the mass ratio of 1:9 to gelatin, stirring for 2h, diluting with phosphate buffered saline to stop the reaction, dialyzing the mixture in deionized water at 40 ℃ for 1 week to remove small molecules and salt, and freeze-drying to obtain the carbon nanotube modified gelatin prepolymer.
(4) Dissolving the gelatin prepolymer modified by the carbon nano tube in phosphate buffered saline with the pH of 7.5 at the dissolving temperature of 70 ℃, fully stirring for 30min, and adding the gelatin prepolymer modified by the carbon nano tube in a mass ratio of 5: 100 of photoinitiator Irgacure 2959, and carrying out ultraviolet B crosslinking reaction for 30min to obtain the carbon nano tube modified gelatin hydrogel. Wherein the irradiation power of the ultraviolet light B is 10mW, and the irradiation wavelength of the ultraviolet light B is 365 nm.
Example 3
(1) To a reaction vessel were added 8mg of cobalt tetra (t-butyl) phthalocyanine, 200mg of ethyl methacrylate monomer, 4.4mg of dimethyl azobisisobutyrate and 1.60mL of chloroform, followed by degassing treatment, and the reaction vessel was sealed. And immersing the sealed reaction container in a constant-temperature water bath at 70 ℃ for reaction for 1h, adding a mixture of acetone and water after the reaction is finished to precipitate the cobalt tetra (tert-butyl) phthalocyanine catalyst, filtering, and performing rotary evaporation to obtain the ethyl methacrylate polymer. The structural formula of the ethyl methacrylate polymer is as follows:
(2) a catalyst layer is deposited on a first substrate and the first substrate is placed in a chemical vapor deposition reactor. Argon gas is introduced, and the temperature is raised to 750 ℃. Then introducing 40% of ethylene, 1% of hydrogen and 59% of nitrogen at the gas flow rate of 3L/min, and reacting for 2min to obtain the carbon nanotube array. A second substrate was taken, and an ethyl methacrylate polymer film having a thickness of 1mm was formed on the second substrate. And in an argon atmosphere, placing the first substrate and the second substrate in a reaction furnace side by side, wherein the first substrate and the second substrate are positioned on the same horizontal plane, and carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate. The irradiation power of the ultraviolet light A is 35mW, the irradiation wavelength of the ultraviolet light A is 196nm, and the irradiation time is 30 min. And after the reaction is finished, closing the ultraviolet light A, and exposing the first substrate to the argon atmosphere until the first substrate is naturally cooled to obtain the modified nanotube array.
(3) Gelatin was dissolved in phosphate buffered saline at pH 7 at a dissolution temperature of 45 ℃. Adding a modified carbon nanotube array with the mass ratio of the modified carbon nanotube array to the gelatin being 1: 13, stirring for 2 hours, and diluting with phosphate buffered saline to stop the reaction. The mixture was then dialyzed in deionized water at 45 ℃ for 1 week to remove small molecules and salts, followed by lyophilization to give a carbon nanotube-modified gelatin prepolymer.
(4) Dissolving the gelatin prepolymer modified by the carbon nano tube in phosphate buffered saline with the pH value of 7, wherein the dissolving temperature is 80 ℃, fully stirring for 30min, adding Irgacure 2959 with the mass ratio of the gelatin prepolymer modified by the carbon nano tube of 8:100, and carrying out ultraviolet B crosslinking reaction for 60min to obtain the gelatin hydrogel modified by the carbon nano tube. Wherein the irradiation power of the ultraviolet light B is 10mW, and the irradiation wavelength of the ultraviolet light B is 365 nm.
Example 4
(1) 7.6mg of bis (borodifluorodiphenylglyoxime) cobalt dihydrate, 190mg of an n-propyl methacrylate monomer, 3.8mg of azobisisoheptonitrile and 1.37mL of chloroform were charged in a reaction vessel, followed by degassing treatment and sealing of the reaction vessel. And (3) immersing the sealed reaction container into a constant-temperature water bath at the temperature of 80 ℃ for reaction for 1 h. After the reaction is finished, adding ethyl acetone to precipitate a cobalt bis (boron difluoro diphenyl glyoxime) dihydrate catalyst, filtering, and performing rotary evaporation to obtain the n-propyl methacrylate polymer. The structural formula of the n-propyl methacrylate polymer is shown below:
(2) a catalyst layer is deposited on a first substrate and the first substrate is placed in a chemical vapor deposition reactor. Introducing helium, and heating to 850 ℃. Then introducing 30% of ethylene, 10% of hydrogen and 60% of nitrogen at the gas flow rate of 1L/min, and reacting for 5min to obtain the carbon nano tube array. A second substrate was taken, and an n-propyl methacrylate polymer film having a thickness of 1mm was formed on the second substrate. And in a helium atmosphere, placing the first substrate and the second substrate in the reaction furnace side by side, wherein the first substrate and the second substrate are positioned on the same horizontal plane, and carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate. The irradiation power of the ultraviolet light A is 35mW, the irradiation wavelength of the ultraviolet light A is 256nm, and the irradiation time is 20 min. And after the reaction is finished, closing the ultraviolet light A, and exposing the first substrate to a helium atmosphere until the first substrate is naturally cooled to obtain the modified nanotube array.
(3) Gelatin was dissolved in phosphate buffered saline at pH 8 at a dissolution temperature of 50 ℃. Adding a modified carbon nanotube array with the mass ratio of the modified carbon nanotube array to the gelatin being 1:19, stirring for 2 hours, and diluting with phosphate buffered saline to stop the reaction. The mixture was then dialyzed in deionized water at 50 ℃ for 1 week to remove small molecules and salts, followed by lyophilization to give a carbon nanotube-modified gelatin prepolymer.
(4) The gelatin prepolymer modified by the carbon nano tube is dissolved in phosphate buffered saline with the pH value of 8, and the dissolving temperature is 80 ℃. And (3) after fully stirring for 30min, adding Irgacure 2959 with the mass ratio of the carbon nanotube modified gelatin prepolymer to the mass ratio of 7: 100, and carrying out ultraviolet B crosslinking reaction for 50min to obtain the carbon nanotube modified gelatin hydrogel. Wherein the irradiation power of the ultraviolet light B is 10mW, and the irradiation wavelength of the ultraviolet light B is 365 nm.
Example 5
(1) To a reaction vessel were added 8.2mg of cobalt tetramethoxyphenylporphyrin, 200mg of methyl methacrylate monomer, 4.4mg of azobisisobutyronitrile and 1.59mL of chloroform, followed by degassing treatment, and the reaction vessel was sealed. And (3) immersing the sealed reaction container into a constant-temperature water bath at 70 ℃ for reaction for 1 h. And after the reaction is finished, adding ethyl acetone to precipitate the catalyst, filtering, and performing rotary evaporation to obtain the methyl methacrylate polymer. The structural formula of the methyl methacrylate polymer is as follows:
(2) a catalyst layer is deposited on a first substrate and the first substrate is placed in a chemical vapor deposition reactor. And introducing nitrogen, heating to 900 ℃, introducing 30% of ethylene, 10% of hydrogen and 60% of nitrogen at the gas flow rate of 2L/min, and reacting for 3min to obtain the carbon nanotube array. A second substrate was taken, and a methyl methacrylate polymer film having a thickness of 1mm was formed on the second substrate. And in a nitrogen atmosphere, placing the first substrate and the second substrate in a reaction furnace side by side, wherein the first substrate and the second substrate are positioned on the same horizontal plane, and carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate. The irradiation power of the ultraviolet light A is 15mW, the irradiation wavelength of the ultraviolet light A is 350nm, and the irradiation time is 30 min. And after the reaction is finished, closing the ultraviolet light A, and exposing the first substrate to a nitrogen atmosphere until the first substrate is naturally cooled to obtain the modified nanotube array.
(3) Gelatin was dissolved in phosphate buffered saline at pH 7.5 at a dissolution temperature of 40 ℃. Adding a modified carbon nanotube array with the mass ratio of the modified carbon nanotube array to the gelatin being 1:19, stirring for 2 hours, and diluting with phosphate buffered saline to stop the reaction. The mixture was then dialyzed in deionized water at 40 ℃ for 1 week to remove small molecules and salts, followed by lyophilization to give a carbon nanotube-modified gelatin prepolymer.
(4) The carbon nanotube modified gelatin prepolymer was dissolved in phosphate buffered saline at pH 7.5 at a dissolution temperature of 70 ℃. And (3) after fully stirring for 30min, adding phenyl-2, 4, 6-trimethyl benzoyl lithium phosphonate with the mass ratio of the carbon nano tube modified gelatin prepolymer to the mass ratio of 5: 100, and carrying out ultraviolet B crosslinking reaction for 30min to obtain the carbon nano tube modified gelatin hydrogel. Wherein the irradiation power of the ultraviolet light B is 15mW, and the irradiation wavelength of the ultraviolet light B is 400 nm.
Comparative example 1
(1) Gelatin was dissolved in phosphate buffered saline at pH 7.5 at a dissolution temperature of 40 ℃. Adding methyl methacrylate with gelatin mass ratio of 1:19, stirring for 2h, diluting with phosphate buffered saline to stop reaction, dialyzing the mixture in deionized water at 40 ℃ for 1 week to remove small molecules and salt, and freeze-drying to obtain methyl methacrylate modified gelatin prepolymer.
(2) The methyl methacrylate-modified gelatin prepolymer was dissolved in phosphate buffered saline at a pH of 7.5 at a dissolution temperature of 70 ℃. And (3) fully stirring for 30min, adding a photoinitiator Irgacure 2959 with the mass ratio of methyl methacrylate modified gelatin prepolymer being 5: 100, and carrying out ultraviolet B crosslinking for 30min to obtain the methyl methacrylate modified gelatin hydrogel. Wherein the irradiation power of the ultraviolet light B is 10mW, and the irradiation wavelength of the ultraviolet light B is 365 nm.
The modified gelatin hydrogels prepared in examples 1 to 5 and comparative example 1 above were tested and the test results were as follows:
(1) polymerization degree test of methacrylate polymer
The methacrylate-based polymers of examples 1 to 5 were measured for molecular weight by gel permeation chromatography and divided by the monomer molecular weight to obtain the degree of polymerization of the methacrylate-based polymers. The results of the experiment are shown in table 1.
TABLE 1 polymerization degree of methacrylate-based polymers
| Sample (I) | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| Degree of polymerization | 3 | 3 | 5 | 8 | 12 |
As can be seen from Table 1, the methacrylate polymers prepared by this method have a degree of polymerization of 3 to 12.
(2) XPS spectrum tests were performed on the raw material pure carbon nanotubes, pure polymethylmethacrylate and the modified carbon nanotubes prepared in example 5 to characterize whether the modified carbon nanotubes prepared in example 5 were grafted with a methyl methacrylate polymer. The test results are shown in FIG. 5-a, FIG. 5-b, FIG. 6-a, FIG. 6-b, FIG. 7-a and FIG. 7-b, respectively.
As can be seen from fig. 7-a and 7-b, the XPS spectra of the modified carbon nanotube array are the superposition of the XPS spectra of the pure carbon nanotubes and the pure polymethyl methacrylate, which indicates that the ultraviolet light treatment can effectively graft the methyl methacrylate polymer onto the carbon nanotubes.
(3) Tensile mechanical property tests were performed on the carbon nanotube-modified gelatin hydrogels prepared in examples 1 to 5 and the non-reinforced methyl methacrylate gelatin hydrogel in comparative example 1, respectively, and a stress-strain diagram was obtained as shown in fig. 8.
Specifically, the tensile strength (MPa) of the above hydrogel was measured by the method of ASTM D-412. The above-mentioned hydrogel was tested for elongation at break (%) by the method of ASTM D-412.
As can be seen from fig. 8, the carbon nano-modified gelatin hydrogel has a significant increase in tensile strength relative to the unreinforced methyl methacrylate gelatin hydrogel. It is shown that the carbon nanotube-modified hydrogels prepared in examples 1 to 5 have good mechanical properties.
(4) The carbon nanotube-modified gelatin hydrogels of examples 1 to 5 and the methyl methacrylate-modified gelatin hydrogel of comparative example 1 were each subjected to a porosity test to characterize the porosity of the hydrogel. When the porosity is tested, firstly, the diameter D and the length L of the hydrogel are tested, the overall volume Vb of the hydrogel is calculated according to the diameter and the length, the skeleton volume Vg of the hydrogel is tested by using a BLP-530 gas porosity instrument of OFI instruments company, and then the formula is adopted: the porosity of the hydrogels of each example and comparative example was calculated as (Vb-Vg)/Vb x 100 (%). The results are shown in Table 2.
TABLE 2
| Sample (I) | Comparative example 1 | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| Porosity of the product | 65％ | 59％ | 55％ | 57％ | 62％ | 60％ |
As can be seen from Table 2, the hydrogel modified by adding carbon nanotubes has a smaller change in porosity than the hydrogel not modified by adding carbon nanotubes. Indicating that the carbon nanotubes do not significantly affect the porosity of the hydrogel.
The above experimental results all show that the carbon nanotube modified gelatin hydrogels prepared in examples 1 to 5 have excellent mechanical properties and have small influence on the porosity of the hydrogel.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A preparation method of gelatin hydrogel modified by carbon nano tubes is characterized by comprising the following steps:
homopolymerization reaction is carried out on methacrylate monomer to obtain the product with the structural formula
The methacrylate polymer of (1), wherein n is 1 to 11, and R ═ CmH2m+1M is 1 to 4;
carrying out grafting reaction on the carbon nanotube array and the methacrylate polymer to obtain a modified carbon nanotube array;
reacting the modified carbon nanotube array with gelatin to obtain a carbon nanotube modified gelatin prepolymer; and
crosslinking the gelatin prepolymer modified by the carbon nano tube to obtain gelatin hydrogel modified by the carbon nano tube;
the method for preparing the methacrylate polymer by homopolymerizing the methacrylate monomer comprises the following steps: carrying out homopolymerization on the methacrylate monomer, a catalyst, an initiator and a solvent under the condition of constant-temperature water bath to obtain the methacrylate polymer, wherein the catalyst is a chelate of divalent cobalt;
the step of carrying out the grafting reaction of the carbon nanotube array and the methacrylate polymer specifically comprises the following steps:
preparing the carbon nanotube array on a first substrate;
depositing the methacrylate-based polymer on a second substrate; and
placing the first substrate and the second substrate on the same horizontal plane side by side, simultaneously carrying out ultraviolet A irradiation treatment on the first substrate and the second substrate under the atmosphere of protective gas so as to enable the gaseous methacrylate polymer to move to the surface of the carbon nanotube array,andthe carbon nano tube array is subjected to grafting reaction, the power of the ultraviolet light A is 15-35 mW, and monochromatic narrow-band light with the irradiation wavelength of 196-350 nm is irradiated;
the step of reacting the modified carbon nanotube array with gelatin specifically comprises:
dissolving the gelatin in phosphate buffered saline to obtain a gelatin solution;
adding the modified carbon nanotube array into the gelatin solution to obtain a mixture; and
dialyzing the mixture, and drying to obtain the carbon nanotube modified gelatin prepolymer;
the step of crosslinking the carbon nanotube modified gelatin prepolymer specifically comprises:
dissolving the carbon nanotube modified gelatin prepolymer in phosphate buffered saline to obtain a carbon nanotube modified gelatin prepolymer solution; and
adding a photoinitiator into the carbon nanotube modified gelatin prepolymer solution, and crosslinking under the irradiation of ultraviolet light B.
2. The method for preparing the carbon nanotube-modified gelatin hydrogel according to claim 1, wherein the mass ratio of the catalyst to the monomer is 4.0-4.2: 100.
3. The method for preparing the carbon nanotube-modified gelatin hydrogel of claim 1, wherein the treatment time of the ultraviolet light A is 20min to 50min, and the distance between the light source of the ultraviolet light A and the first substrate and the second substrate is 2mm to 20 mm.
4. The method for preparing a carbon nanotube-modified gelatin hydrogel according to claim 1, wherein the temperature during the dialysis is 40 to 50 ℃.
5. The method for preparing the carbon nanotube-modified gelatin hydrogel according to claim 1, wherein the mass ratio of the modified carbon nanotube array to the gelatin is 1:9 to 1: 19.
6. The method for preparing the carbon nanotube-modified gelatin hydrogel according to claim 1, wherein the photoinitiator is a basf photoinitiator Irgacure 2959 or lithium phenyl (2,4, 6-trimethylbenzoyl) phosphate.
7. The method for preparing the carbon nanotube-modified gelatin hydrogel according to claim 1 or 6, wherein the mass ratio of the photoinitiator to the carbon nanotube-modified gelatin prepolymer is 5-8: 100.
8. The method for preparing the carbon nanotube-modified gelatin hydrogel according to claim 1 or 6, wherein the ultraviolet light B is monochromatic narrow-band light with a wavelength of 365-400 nm, the irradiation power of the ultraviolet light B is 10-15 mW, and the treatment time of the ultraviolet light B is 30-60 min.
9. A carbon nanotube-modified gelatin hydrogel prepared by the method for preparing a carbon nanotube-modified gelatin hydrogel according to any one of claims 1 to 8.
10. Use of the carbon nanotube-modified gelatin hydrogel of claim 9 in the preparation of a three-dimensional cell culture scaffold or in the preparation of a biomimetic extracellular matrix.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811578203.3A CN109749096B (en) | 2018-12-21 | 2018-12-21 | Carbon nanotube modified gelatin hydrogel and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811578203.3A CN109749096B (en) | 2018-12-21 | 2018-12-21 | Carbon nanotube modified gelatin hydrogel and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109749096A CN109749096A (en) | 2019-05-14 |
| CN109749096B true CN109749096B (en) | 2021-06-15 |
Family
ID=66403933
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811578203.3A Active CN109749096B (en) | 2018-12-21 | 2018-12-21 | Carbon nanotube modified gelatin hydrogel and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109749096B (en) |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5587431A (en) * | 1995-04-21 | 1996-12-24 | E. I. Du Pont De Nemours And Company | Synthesis of terminally unsaturated oligomers |
| CN101693125B (en) * | 2009-10-12 | 2013-06-19 | 北京科技大学 | Process for preparing biocompatible directional carbon nanotube array reinforced composite hydrogel |
| CN105107019B (en) * | 2015-09-10 | 2018-05-18 | 西南交通大学 | A kind of preparation method of the high-strength hydrogel of infrared response for articular cartilage reparation |
| US10799618B2 (en) * | 2016-01-21 | 2020-10-13 | Saint Louis University | Methods of transferring carbon nanotubes on a hydrogel |
| CN107236135A (en) * | 2017-07-07 | 2017-10-10 | 中国科学院理化技术研究所 | A kind of gelatin hydrogel and its preparation method and application |
| CN108314009B (en) * | 2018-03-30 | 2020-11-20 | 烯湾科城(广州)新材料有限公司 | Surface modification method of carbon nanotube array |
-
2018
- 2018-12-21 CN CN201811578203.3A patent/CN109749096B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109749096A (en) | 2019-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhou et al. | Surface functionalization of biomaterials by radical polymerization | |
| Sakellariou et al. | Surface-initiated polymerization from carbon nanotubes: strategies and perspectives | |
| Roy et al. | Synthesis of natural–synthetic hybrid materials from cellulose via the RAFT process | |
| Kakwere et al. | Design of complex polymeric architectures and nanostructured materials/hybrids by living radical polymerization of hydroxylated monomers | |
| Oktay et al. | Immobilization of α-amylase onto poly (glycidyl methacrylate) grafted electrospun fibers by ATRP | |
| CN113444264B (en) | Preparation method and application method of double-network hydrogel for three-dimensional cell culture | |
| KR20110062804A (en) | Interpenetrating polymer network hydrogel and preparation method thereof | |
| Ata et al. | Polymer nano-hybrid material based on graphene oxide/POSS via surface initiated atom transfer radical polymerization (SI-ATRP): Its application in specialty hydrogel system | |
| Xu et al. | Thermoresponsive poly (N-isopropyl acrylamide)-grafted polycaprolactone films with surface immobilization of collagen | |
| CN114106227B (en) | Preparation method of structured hydrogel and hydrogel heart and valve | |
| Yin et al. | Argon plasma-induced graft polymerization of PEGMA on chitosan membrane surface for cell adhesion improvement | |
| Hou et al. | Surface nanostructures based on assemblies of polymer brushes | |
| CN109749096B (en) | Carbon nanotube modified gelatin hydrogel and preparation method and application thereof | |
| TWI330657B (en) | Super-low fouling carboxybetaine materials and related methods | |
| CN113975462A (en) | Preparation method of dopamine-functionalized neuroactive hydrogel | |
| CN110669168A (en) | Modifying agent, preparation method and use method thereof and medical material | |
| CN106317714B (en) | The low-temperature plasma modified processing method of nano-aluminium oxide | |
| CN109758608B (en) | Printable composite hydrogel with high toughness, preparation method and application | |
| CN101602835A (en) | A kind of preparation method of carbon mano-tube composite of polymer graft | |
| Mittal et al. | Functionalized polystyrene latex particles as substrates for ATRP: Surface and colloidal characterization | |
| JP6156874B2 (en) | Bioinert film, coating liquid, production method thereof, and bioinert-treated substrate | |
| Jayasinghe et al. | A soft lithography method to generate arrays of microstructures onto hydrogel surfaces | |
| WO2012029731A1 (en) | Method for manufacturing cell culture substrate | |
| CN115073770A (en) | Physical gel based on zwitterion modified polysaccharide and preparation method thereof | |
| JP2018145216A (en) | Enzymatic synthesis of three-dimensional structure comprising cellulose oligomer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |